The present study investigates the effects of temporal synergism of 5-hydroxytryptophan (5-HTP) and L-dihydroxyphenylalanine (L-DOPA) and pineal function (pinealectomy and melatonin administration) on gonadal response and nitric oxide (NO) activity in Japanese quail. Three-week-old male quails were divided into 9 groups, as follows: (I) control (treated with normal saline); (II) 8-h and (III) 12-h (receiving L-DOPA after the administration of 5-HTP [5mg/100g body weight] at the interval of 8 and 12h, respectively, over a period of 13 days); (IV) SO (sham operated); (V) Px (pinealectomized); (VI) Veh (vehicle treated control); (VII) Mel (melatonin 25 microg/100g body weight for 45 days); (VIII) 12-h+Veh; and (IX) 12-h+Mel (same as in (III) but further treated with the vehicle or melatonin, respectively, for 32 days). Body weight and cloacal gland volume were recorded weekly, whereas spermatogenesis, plasma testosterone concentration, and total nitrite and nitrate concentration in plasma, hypothalamus, and testes were measured at the termination of the study. Results indicate that compared to the controls, 8-h, Mel, and 12-h+Mel treatments suppressed whereas 12-h and Px stimulated the reproductive system. Further, a significant increase in plasma testosterone, total nitrite, and nitrate in the 12-h and Px groups and a decrease in these concentrations in 8-h, Mel and 12-h+Mel quail were observed. It is evident that 5-HTP and L-DOPA given at the interval of 8h, as well as melatonin, are potent inhibitors of gonadal development, and the gonado stimulatory effect of the 12-h temporal relationship of the neurotransmitter precursors is masked by the inhibitory effect of melatonin. It is concluded that modulation of the daily phase relationship of neural oscillations and pineal function alters gonadal function and NO activity. Further, NO exhibits a parallel relationship with gonadal function in Japanese quail; this relationship appears to be mediated through NO activity, suggesting a causal relationship, although the mechanism between the 2 systems remains a topic of discussion.